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MOLECULAR BIOLOGY > 3. REAGENT PREPARATION - LAB > Flashcards

3. REAGENT PREPARATION - LAB Flashcards

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1
Q

Where can you find instructions for preparing reagents and quantities used?

A

Written laboratory protocol

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2
Q

What should be documented and matched to test performance in amplification methods?

A

Lot numbers and working stocks of probes and primers

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3
Q

Why should probe sequences used for a given test be recorded?

A

Because probes for linkage analysis and array technology are periodically updated

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4
Q

What is the purpose of an RNA primer in vivo?

A

Initiates DNA replication

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5
Q

What is the purpose of RNA primers in vitro (synthetically made primers)?

A

Used in DNA amplification

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6
Q

What is a critical component of PCR procedures?

A

Primers

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7
Q

What is the lyophilized form of a primer?

A

freeze dried or powdered form

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8
Q

What information is included in the primer product?

A

Quality, Method of purification, Molecular weight, Number of micrograms of dried primers

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9
Q

What is the typical length of oligonucleotide primers in base pairs?

A

18-25 or 18-30

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10
Q

What do probes bind to directly?

A

Target sequence

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11
Q

What is the purpose of probes?

A

Detection of target sequence or hybridization techniques

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12
Q

In what forms are probes supplied?

A

Solution or dried form

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13
Q

How should new working stocks of probes be treated?

A

As new reagent lots

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14
Q

What needs to be done to dried form probes or primers?

A

Dissolve in solvent to make stock solution

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15
Q

What should be done with stock solution to make a working solution?

A

Dilute it

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16
Q

What are analyte-specific reagents (ASRs)?

A

Probes, primers, antibodies

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17
Q

What does analyte-specific reagents detect?

A

Presence or absence of a particular substance

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18
Q

Where are analyte-specific reagents used?

A

Tissue typing, infectious disease, PCR

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19
Q

What are in vitro diagnostic (IVD) devices?

A

Instruments or systems for diagnosis

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20
Q

What are IVD reagent sets used for?

A

Collecting, preparing, or examining specimens

21
Q

How are IVD devices classified?

A

CLASS I, CLASS II, CLASS III

22
Q

Reagent categories

A

ASRs
IVD
IVAT
RUO IUO

23
Q

This category is intended to accommodate the use of promising high complexity technologies facing long clearance processes

A

In vitro analytical test (IVAT) reagents

24
Q

are not intended for diagnostic use

A

Research Use Only (RUO)

25
Q

can be used in patient samples with proper institutional review and informed consent

A

Investigational Use Only (IUO)

26
Q

reagents are used to gather data that may result in the advancement of a product’s status

A

Research Use Only (RUO) and Investigational Use Only (IUO) reagents

27
Q

What is the first step in making reagents and buffers?

A

Go through the protocol for the experiment and write down all the solutions used.

28
Q

What should you decide about the solution concentration?

A

Decide whether to make diluted or concentrated solutions.

29
Q

What should you check regarding chemical availability?

A

Find out if the chemicals you need are available in the lab.

30
Q

What should you check for safety information?

A

MSDS (Material Safety Data Sheet)

31
Q

What label should you look for on hazardous materials?

A

Hazardous material classification label

32
Q

Where should you read for warnings about the reagent?

A

Reagent bottle/container

33
Q

What should you do if there are no warnings on the label?

A

Look up the material in the Merck Index

34
Q

What if the label is in another language or you can’t find a listing?

A

Consult the manufacturer

35
Q

What is a concern during the analytical phase of work?

A

Exposure to various hazardous reagents

36
Q

What neurotoxin is used for protein and sequencing gels?

A

Acrylamide

37
Q

What mutagen intercalates in DNA and is used to label nucleic acids?

A

Ethidium bromide (EtBr)

38
Q

What highly corrosive substance can burn skin and should be used in a fume hood?

A

Phenol

39
Q

What reagent, used to inhibit proteinases, can be fatal if swallowed or absorbed through the skin?

A

Phenylmethyl-sulfonate fluoride (PMSF)

40
Q

What detergent is a burning powder and extremely irritating to nasal passages?

A

Sodium dodecyl sulfate (SDS)

41
Q

What method is used for sterilizing most buffers?

A

Autoclave

42
Q

What material are tips and microcentrifuge tubes made of for autoclaving?

A

High-density polyethylene

43
Q

What should be avoided when autoclaving tips and microcentrifuge tubes?

A

Proteins

44
Q

What should you check for in solutions before autoclaving?

A

Acidity or alkalinity

45
Q

What substances should not be autoclaved?

A

Corrosives, solvents, volatiles, liquids with bleach/formalin/glutaraldehyde, buffers with detergents (e.g., 10% SDS), heat-labile ingredients, DTT, and β-mercaptoethanol

46
Q

Storage of buffers in room temperature

A

Detergents
Ethanol
Buffers
Concentrated solutions
Acid and bases

47
Q

Storage of buffers and solutions in 4C

A

Bacterial cultures
Buffers
DNA
Media
PCR products
Serum

48
Q

Storage of buffer and solutions in -20c

A

Enzymes
RNA

49
Q

Storage of buffer and solutions in -70c

A

Frozen bacterial cultures
Lipids

MOLECULAR BIOLOGY (7 decks)

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